Abstract

We examined the direct effects of toxaphene and endrin, chlorinated insecticides that are widespread in the environment, on myometrial contractions and on the secretion of hormones involved in regulating these contractions. Granulosa, luteal, endometrial and myometrial cells, and myometrial strips from non-pregnant cows were incubated with both insecticides at environmentally relevant doses.

Toxaphene inhibited and endrin stimulated the secretion of testosterone and oestradiol from granulosa cells. Toxaphene also inhibited and endrin stimulated the expression of the mRNA encoding the precursor of oxytocin (OT), as well OT secretion in luteal cell cultures. Moreover, endrin increased OT secretion from granulosa cells. Neither insecticide exerted an effect on progesterone secretion from luteal cells. Only toxaphene decreased the secretion of prostaglandins (PGF2 and PGE2) from endometrial cells. Meanwhile, only endrin decreased basal myometrial contractions, which was accompanied by inhibition of PGF2 secretion from the myometrium. Both endrin and toxaphene also decreased the force of the OT-stimulated myometrial contractions, whereas only toxaphene inhibited the stimulatory effect of OT on the force of myometrial contractions.

In contrast to endrin, toxaphene decreased synthesis and secretion of one of the primary stimulators of myometrial contractions (OT) and indirectly inhibited OT signal reception in the myometrium by reducing E2 secretion. Both insecticides decreased OT-stimulated myometrial contractions; therefore, they may inhibit further transmission of the OT signal. Moreover, endrin inhibited basal myometrial contractions, potentially resulting from reduced PGF2 secretion from the myometrium. Our data indicate the potential of these insecticides to disturb the course of the oestrous cycle or fertilisation.

Abstract

Purpose

Ocular autonomic control is mediated by sympathetic and parasympathetic nerve fibres. Their interactions are complemented by primary afferent nerve fibers of and intrinsic choroidal neurons (ICN). As the vasodilatative neuropeptide, vasoactive intestinal peptide (VIP), is expressed in extrinsic and intrinsic ocular neurons, it is of special interest in ophthalmic research. Since circadian changes of ocular blood flow are known in humans and birds, this study aimed at investigating VIP expression at different daytimes in chicken choroid, the preferred model species in ICN research.

Methods

12 eyes of 12 chickens were retrieved, slaughtered at 8.00-9.30 a.m. (n=6) and 8.00 p.m. (n=6), respectively, and choroidal wholemounts were prepared for immunofluorescence of VIP. VIP-positive ICN of both groups were quantified and density of VIP-positive axons assessed semi-quantitatively. In 28 additional eyes retrieved in the morning (n=14) and evening (n=14), choroidal VIP content was determined by ELISA. Morning and evening data were analyzed statistically.

Results

(1) Numbers of VIP positive neurons differed significantly between morning: (239.17±113.9) and evening: (550.83±245.7; p=0.018). (2) Numbers of VIP-positive perikarya were significantly more accumulated in the temporal part of the choroid in the evening than in the morning (p=0.026). (3) VIP positive axon density was found to be similar throughout the choroid in the morning and evening. (4) ELISA demonstrated a significant difference of VIP content (p=0.012) in tissues harvested in the morning (145.41±43.3?pg/ml) compared to evening (221.44±106.3?pg/ml).

Conclusions

As VIP positive axon density was similar in the morning and the evening throughout the choroid, PPG and ICN seemed to contribute equally to the axon network. Yet, changes in the total choroidal VIP content, the numbers of VIP positive perikarya, reflecting the intracellular VIP content, and their topographical distribution at two different days-times argue for a different status of activation of both neuronal sources. The higher VIP content in the evening, compared to the morning, correlates with a known circadian rhythm of a lower IOP and a higher choroidal thickness at night. Thus, these changes may argue for a potential role of ICN in the regulation of ocular homeostasis and integrity.

Abstract

Aim

The aim of the study was to determine HMGB-1 concentration in serum and cerebrospinal fluid (CSF) of patients suffering from tick-borne encephalitis (TBE) and neuroborreliosis (NB). We focused on HMGB-1 measurement in CSF or sera in order to establish whether it could help to differentiate between NB and TBE.

Methods

80 patients with meningitis and meningoencephalitis were enrolled in the study. Patients were divided into two groups: Group I - patients with NB (n = 40) and Group II - patients with TBE (n = 40). Diagnosis was made basing on the clinical picture, CSF examination and specific antibodies presence in serum and CSF. The control group for the evaluation of the parameters in serum were healthy blood donors (n = 25), while control group for evaluation of CSF consisted of patients with excluded CNS inflammatory process. Concentrations of HMGB-1 was measured by ELISA method using commercial kit [HMGB-1 Elisa Kit, (EIAab, China)]. The results were statistically analyzed using STATISTICA 10, Gretl, ROC, Pearson correlation coefficient.

Results and Conclusions

HMGB-1 is associated with the development of inflammatory process in the CNS caused by both tick-borne pathogens: viral (TBE) and bacterial (Lyme borreliosis). Measurement of serum HMGB-1 concentration in the early stages of both diseases of CNS may contribute to the differentiation between TBE and NB, which may have clinical impact for patients bitten by ticks.

Abstract

Methylmalonic aciduria (MMA) is a disorder of organic acid metabolism resulting from a functional defect of the mitochondrial enzyme, methylmalonyl-CoA mutase (MCM). The main treatments for MMA patients are dietary restriction of propiogenic amino acids and carnitine supplementation. Liver or combined liver/kidney transplantation has been used to treat those with the most severe clinical manifestations. Thus, therapies are necessary to help improve quality of life and prevent liver, renal and neurological complications. Previously, we successfully used the TAT-MTS-Protein approach for replacing a number of mitochondrial-mutated proteins. In this targeted system, TAT, an 11 a.a peptide, which rapidly and efficiently can cross biological membranes, is fused to a mitochondrial targeting sequence (MTS), followed by the mitochondrial mature protein which sends the protein into the mitochondria. In the mitochondria, the TAT-MTS is cleaved off and the native protein integrates into its natural complexes and is fully functional. In this study, we used heterologous MTSs of human, nuclear-encoded mitochondrial proteins, to target the human MCM protein into the mitochondria. All fusion proteins reached the mitochondria and successfully underwent processing. Treatment of MMA patient fibroblasts with these fusion proteins restored mitochondrial activity such as ATP production, mitochondrial membrane potential and oxygen consumption, indicating the importance of mitochondrial function in this disease. Treatment with the fusion proteins enhanced cell viability and most importantly reduced MMA levels. Treatment also enhanced albumin and urea secretion in a CRISPR/Cas9-engineered HepG2 MUT (-/-) liver cell line. Therefore, we suggest using this TAT-MTS-Protein approach for the treatment of MMA.

Abstract

Parkinson's disease (PD) is a progressive neurodegenerative disorder with behavioral and motor abnormalities. Androst-5-ene-3beta, 17beta-diol (ADIOL), an estrogen receptor (ER)beta agonist, was found to mediate a transrepressive mechanism that selectively modulates the extent of neuroinflammation and, in turn, neurodegeneration. In consensus, ER beta polymorphism was more frequently detected in early‐onset PD patients. Thus, in an approach to elucidate the role of ER beta agonists on PD, our study was designed to investigate the possible neuroprotective effect of ADIOL, in three dose levels (0.35, 3.5, 35?mg/kg/day), against rotenone (ROT)-induced PD rat model. Amelioration in striatal dopamine (DA), nuclear factor-kappa B (NF-kB), and the expression of down-stream inflammatory mediators, as well as apoptotic markers were observed in the striatum and substantia nigra (SN) upon pre-treatment with the three doses of ADIOL. Similarly, light microscopy (LM) examination revealed declined degeneration of neurons upon pretreatment with ADIOL. Significant improvement in nigral tyrosine hydroxylase (TH) and reduction of nigral alpha-synuclein densities were also detected after ADIOL pre-treatment with better results frequently achieved with the middle dose (3.5?mg/kg/day). The middle dose of ADIOL showed behavioral improvement, with elevation in the ATP level, which was emphasized by the improvement in mitochondrial integrity observed upon electron microscopy (EM) examination. In conclusion, the present study confirmed for the first time the ability of ADIOL to protect against neuroinflammation and, in turn, neurodegeneration process and motor dysfunction in PD animal model, which was more obviously observed with the middle dose.